A Comprehensive Panel of Turn‐On Caspase Biosensors for Investigating Caspase Specificity and Caspase Activation Pathways

Shekhawat, Sujan S.; Campbell, Sean T.; Ghosh, Indraneel

doi:10.1002/cbic.201100372

Cited by 9 publications

(9 citation statements)

References 69 publications

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“…) and that recombinant caspase‐4 can process procaspase‐3 into p20 and p10 subunits (Shekhawat et al . ). Autoprocessing of caspase‐5 (Viganò et al .…”

Section: Resultsmentioning

confidence: 97%

Proplatelet formation in megakaryocytes is associated with endoplasmic reticulum stress

Morishima

Nakanishi

2016

Genes to Cells

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Although previous studies suggest that proplatelet formation in megakaryocytes involves caspase-3, the mechanism underlying the activation of caspase-3 is unknown. Here, we analyzed caspase activation in a human megakaryoblastic cell line, MEG-01, which forms proplatelets spontaneously. Specific activation of caspase-3 and caspase-4 was found in proplatelets. Consistent with previous observations of caspase-4 autoactivation in response to endoplasmic reticulum (ER) stress, several ER stress marker proteins were expressed during proplatelet formation. A pharmacological ER stressor enhanced platelet production via proplatelet formation, whereas inhibition of caspase-4 caused suppression. These results suggest that ER stress is a mechanism underlying the maturation of megakaryocytes.

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“…) and that recombinant caspase‐4 can process procaspase‐3 into p20 and p10 subunits (Shekhawat et al . ). Autoprocessing of caspase‐5 (Viganò et al .…”

Section: Resultsmentioning

confidence: 97%

Proplatelet formation in megakaryocytes is associated with endoplasmic reticulum stress

Morishima

Nakanishi

2016

Genes to Cells

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“…The modularity of this method has allowed other split-protein type enzymes, such as β-lactamase, to be used as the enzyme amplifier and this signal amplification method has also been applied to the detection of caspases by simply changing the enzyme-cleavable linkage between the coiled-coil and the split-protein. 210 Utilising enzyme cascades for signal amplification relies on the ability to structurally modify enzymes to restrict enzyme activity without denaturisation yet in the presence of the target, enzyme activity must be easily restored. Although a number of examples have been demonstrated, this still remains a significant challenge.…”

Section: Double-catalyst Signal Amplificationmentioning

confidence: 99%

Approaches towards molecular amplification for sensing

Goggins

Frost

2016

Analyst

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Diagnostic assays that rely on molecular interactions have come a long way; from initial reversible detection systems towards irreversible reaction indicator-based methods. More recently, the emergence of innovative molecular amplification methodologies has revolutionised sensing, allowing diagnostic assays to achieve ultra-low limits of detection. There have been a significant number of molecular amplification approaches developed over recent years to accommodate the wide variety of analytes that require sensitive detection. To celebrate this achievement, this comprehensive critical review has been compiled to give a broad overview of the many different approaches used to attain amplification in sensing with an aim to inspire the next generation of diagnostic assays looking to achieve the ultimate detection limit. This review has been created with the focus on how each conceptually unique molecular amplification methodology achieves amplification, not just its sensitivity, while highlighting any key processes. Excluded are any references that were not found to contain an obvious molecular amplifier or amplification component, or that did not use an appropriate signal readout that could be incorporated into a sensing application. Additionally, methodologies where amplification is achieved through advances in instrumentation are also excluded. Depending upon the type of approach employed, amplification strategies are divided into four categories: target, label, signal or receptor amplification. More recent, more complex protocols combine a number of approaches and are therefore categorised by which amplification component described within was considered as the biggest advancement. The advantages and disadvantages of each methodology are discussed along with any limits of detection, if stated in the original article. Any subsequent use of the methodology within sensing or any other application is also mentioned to draw attention to its practicality. The importance of amplification within sensing is wholly emphasised while perspectives on the future direction of the field are also shared.

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“…If it is too short, it cannot effectively separate the two luciferase domains spatially causing high background luminescent signal. To circumvent this limitation, autoinhibited coiled coil strategies (two fragments of firefly luciferase are attached to two different coiled coil partners) were proposed for reduction of background. This made it possible to increase the intensity of bioluminescent signals 1000‐fold.…”

Section: Novel Genetically Encoded Biosensors Based On Modified Firefmentioning

confidence: 99%

“…It must be noted that the main indicator of the efficiency of the fusion protein synthesis is preservation of properties of both domains in the fusion structure and high yield of the target protein. Development of fusion proteins for BRET systems as well as based on split luciferases is also one of the hot topics.…”

Section: Introductionmentioning

confidence: 99%

Firefly Luciferase‐based Fusion Proteins and their Applications in Bioanalysis

Smirnova

Ugarova

2016

Photochem & Photobiology

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Firefly luciferase is widely used in molecular biology and bioanalytical systems as a reporter molecule due to the high quantum yield of the bioluminescence, availability of stable mutant forms of the enzyme with prescribed spectral characteristics and abundance of bacterial expression systems suitable for production of recombinant proteins in limitless quantities. In this review, we described fusion proteins of luciferase with biotin-binding domain and streptavidin, with proteins A and G, antibodies, with DNA-and RNA-binding proteins, as well as fusion proteins designed for BRET systems. The firefly luciferase-based fusion proteins are represented as an effective tool for the development of different bioanalytical systems such as (1) systems in which luciferase is attached to the surface of the target and the bioluminescence signal is detected from the specific complexes formed; (2) BRET-based systems, in which the specific interaction induces changes in the bioluminescence spectrum; and (3) systems that use modified or split luciferases, in which the luciferase activity changes under the action of the analyte. All these systems have wide application in biochemical analysis of physiologically important compounds, for the detection of pathogenic bacteria and viruses, for evaluation of proteinprotein interactions, assaying of metabolites involved in cell communication and cell signaling.Abbreviations: Ab, antibody; b, biotin; bccp, biotin carboxyl carrier protein domain of acetyl-CoA carboxylase from E. coli; HRP, horseradish peroxidase; IA, immunoassay; KPBT, Klebsiella pneumoniae oxaloacetate decarboxylase; Luc, firefly luciferase; PG, pepsinogen; PSA, prostate-specific antigen; SA, streptavidin.

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A Comprehensive Panel of Turn‐On Caspase Biosensors for Investigating Caspase Specificity and Caspase Activation Pathways

Cited by 9 publications

References 69 publications

Proplatelet formation in megakaryocytes is associated with endoplasmic reticulum stress

Proplatelet formation in megakaryocytes is associated with endoplasmic reticulum stress

Approaches towards molecular amplification for sensing

Firefly Luciferase‐based Fusion Proteins and their Applications in Bioanalysis

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